AMBERCHROM CG300 Chromatographic Grade Resin For Chromatographic Purification Description AMBERCHROM chromatographic media are macroporous, polymeric resins useful for adsorption and reversed phase liquid chromatography. They are designed for laboratory and process scale purifications of proteins, peptides, nucleic acids, antibiotics, and small molecular weight pharmaceuticals. AMBERCHROM CG300 chromatographic grade resin is an insoluble polystyrene divinylbenzene polymer manufactured for high value chromatographic applications. Its high surface area, unique pore size and pore volume distribution make it ideally suited for separation of peptides. AMBERCHROM CG300 has high capacity for many pharmaceutical compounds and has been commercially proven for many years. It is an excellent technical and economical alternative to RPC silica, and can be used in high resolution, low pressure chromatography. AMBERCHROM CG300 is suitable for use in many pharmaceutical applications in the front end capture, purification, and desalting modes of operation depending on the particle size selected. AMBERCHROM CG300 is available in three different particle size ranges (35, 75, and 120 microns), and is supplied as a slurry in 20% ethanol. AMBERCHROM CG300 is ideally suited for operation with the entire ph range, and can be easily cleaned in place (CIP) with most organic solvents and dilute acids and bases. Regulatory Status A Material Regulatory Support (MRS) package is maintained for AMBERCHROM CG300 resin. It is available upon request under CDA for users of this product. This material is manufactured under strict controls, and plant audits by potential customers are welcomed. Clean in Place Unlike RPC silica, AMBERCHROM CG300, due to its polymeric nature and lack of bonded phase, can be cleaned in place (CIP) with most organic solvents and low concentrations of acids and bases. As the graph below demonstrates, there is no loss in capacity for Insulin after exposure to 0.5 M NaOH for 180 days at ambient temperature or 100 days exposure at 60 C.
In addition to the excellent chemical resistance of AMBERCHROM CG300, it also exhibits low swelling in common solvents as shown below: Customer Application: Decapeptide Mixture AMBERCHROM CG300S was used to separate a mixture of decapeptides that varied by only one amino acid. The adsorbent which was loaded to 15-20% of the total capacity demonstrated Langmuir behavior and gave an excellent separation of this mixture. Synthetic Peptide Purification S2: Ac-Arg-Gly-Gly-Gly-Gly-Leu-Gly-Leu-Gly-Lys-amide (10%) S3: Ac-Arg-Gly-Ala-Gly-Gly-Leu-Gly-Leu-Gly-Lys-amide (80%) S4: Ac-Arg-Gly-Val-Gly-Gly-Leu-Gly-Leu-Gly-Lys-amide (10%) *Ac = N-acetyl, amide = C-amide
IR Spectrum of Amberchrom CG300 Importance of Selectivity This example shows the separation of insulin from a well known impurity, desamido insulin. The graph compares RPC silica to AMBERCHROM CG300. These two adsorbents have the same number of theoretical plates, so the difference in behavior is related to polymer selectivity.
Typical Physical Properties These properties are typical but do not constitute specifications. Matrix Functional Groups Physical form Shipping solvent Polystyrene/divinylbenzene adsorbent None Opaque white beads 20% ethanol Surface area 700 m 2 /g Pore size [1] 300 Å Mean Diameter S grade: 35 microns M grade: 75 microns C grade: 120 microns Uniformity coefficient 1.7 Chemical resistance Insoluble in dilute solutions of acids or bases and common solvents: IPA, ACN, MeOH. [1] Test methods are available on request. Selected References Characterization of a family of polymeric resins with average pore diameters of 150, 300, and 1000 for the preparative reverse phase purification of polypeptides ; Cartier, Peter G.; Deissler, Karl C.; Maikner, John J.; Kraus, Michael ; Spec. Publ. - R. Soc. Chem. (1994), 158 (Separations for Biotechnology 3), 100-5 CODEN: SROCDO; ISSN: 0260-6291. Validation of a theoretical model for adsorption using cephalosporin C and polymeric reversed-phase resins ; Firouztale', E.; Maikner, J. J.; Deissler, K. C.; Cartier, P. G. ; J. Chromatogr. (1994), 658(2), 361-70 CODEN: JOCRAM; ISSN: 0021-9673. The utility of polymeric reversed phase packings for the purification of peptides, proteins and antibiotics ; Cartier, P. G.; Deissler, K. C.; Maikner, J. J. ; Sep. Biotechnol. 2, [Pap. Int. Symp.], 2nd (1990), 275-84. Editor(s): Pyle, D. L. Publisher: Elsevier, London, UK. CODEN: 56YXAO. Use of resins for trichothecene production in liquid cultures ; Jarvis, Bruce B.; Armstrong, Catherine A.; Zeng, Ming ; J. Antibiot. (1991), 44(3), C1 CODEN: JANTAJ; ISSN: 0021-8820. Analytical- and preparative-scale chromato-graphic separation of phenylalanine from aspartame using a new polymer sorbent ; Ladisch, Michael R.; Hendrickson, Richard L.; Firouztale, Edward ; J. Chromatogr. (1991), 540(1-2), 85-101 CODEN: JOCRAM; ISSN: 0021-9673. Use of resins for trichothecene production in liquid cultures ; Jarvis, Bruce B.; Armstrong, Catherine A.; Zeng, Ming ; J. Antibiot. (1990), 43(11), 1502-4 CODEN: JANTAJ; ISSN: 0021-8820. Automated purification and quantification of oligonucleotides ; Ivanetich, K. M.; Reid, R. C.; Ellison, R.; Perry, K.; Taylor, R.; Reschenberg, M.; Mainieri, A.; Zhu, D.; Argo, J.; Cass, D.; Strickland, C. ; BioTechniques (1999), 27(4), 810,812,814-818,820,822-823 CODEN: BTNQDO; ISSN: 0736-6205. Purification of LL-AO341 antibiotics derived from fermentation broth by large scale preparative reversed phase chromatography on Amberchrom CG161 M.; Williams, David R.; Lee, M. D.; Pinho, F.; Borders, D. B. ; Abstracts of Papers American Chemical Society, (1994) Vol. 207, No. 1-2, pp. BTEC 89. ; Meeting Info.: 207th National Meeting of the American Chemical Society San Diego, California, USA March 13-17, 1994 ;ISSN: 0065-7727. Purification of fermentation products with polymeric media ; Cartier, P. G. ; Maikner, J. (1); Deissler, K. C.; Firouztale, E. Abstracts of Papers American Chemical Society, (1993) Vol. 205, No. 1-2,pp. BIOT 18. ; Meeting Info.: 205th ACS (American Chemical Society) National Meeting Denver, Colorado, USA March 28-
April 2, 1993 ISSN: 0065-7727. Process Economics Aspects of RPC Purification; Kinzey, M.; Kraus, M.; Fisher, J.; Maikner, J.; Rosen, R.; Recovery of Biological Products IX; May 23-28, 1999; Whistler, CA. Process Development and Scale up Using Polymeric RPC Resins; Kinzey, M.; Fisher, J.; O'Donnell, J.K.; Rosen, R.E.; Maikner. J.J.; Kraus, M/M., Iuliano, S.J.; Prep 98; May 31-June 3, 1998, Washington, DC. Strategies for Optimizing Peptide Purifications Using Amberchrom CG300S; Kinzey, M.; Deissler, K.; Fisher, J.; Maikner, J.; Kraus, M.; Rosen, R.; European Forum on Advances in Industrial Downstream Processing; May 13-15, 1998, Stuttgart, Germany. Process Economics Aspects of RPC Purification; Kinzey, M.; Kraus, M.; Fisher, J.; Maikner, J.; Rosen, R.; 3rd HIC/RPC Bioseparations Conference; February 5-9, 2001; Athens, Greece. Ordering Information
AMBERCHROM is a trademark of Rohm and Haas Company, Philadelphia, U.S.A. Ion exchange resins and polymeric adsorbents, as produced, contain by-products resulting from the manufacturing process. The user must determine the extent to which organic by-products must be removed for any particular use and establish techniques to assure that the appropriate level of purity is achieved for that use. The user must ensure compliance with all prudent safety standards and regulatory requirements governing the application. Except where specifically otherwise stated, Rohm and Haas Company does not recommend its ion exchange resins or polymeric adsorbents, as supplied, as being suitable or appropriately pure for any particular use. Consult your Rohm and Haas technical representative for further information. Acidic and basic regenerant solutions are corrosive and should be handled in a manner that will prevent eye and skin contact. Nitric acid and other strong oxidising agents can cause explosive type reactions when mixed with ion exchange resins. Proper design of process equipment to prevent rapid buildup of pressure is necessary if use of an oxidising agent such as nitric acid is contemplated. Before using strong oxidising agents in contact with ion exchange resins, consult sources knowledgeable in the handling of these materials. Rohm and Haas Company makes no warranties either expressed or implied as to the accuracy or appropriateness of this data and expressly excludes any liability upon Rohm and Haas arising out of its use. We recommend that the prospective users determine for themselves the suitability of Rohm and Haas materials and suggestions for any use prior to their adoption. Suggestions for uses of our products of the inclusion of descriptive material from patents and the citation of specific patents in this publication should not be understood as recommending the use of our products in violation of any patent or as permission or license to use any patents of the Rohm and Haas Company. Material Safety Data Sheets outlining the hazards and handling methods for our products are available on request. Rohm and Haas, 2006 All rights reserved. February 2005